Polypharmacophoric agents

ABSTRACT

One aspect of the present invention relates to polypharmacophoric compounds. In certain embodiments, the polyphamacophore compounds comprise individual pharmacophore units selected from the group consisting of D-1 agonists, D-2 agonists, D-3 agonists, D-4 agonists, irreversible monoamine inhibitors, reversible monoamine inhibitors, monoamine transporter inhibitors, COMT inhibitors, MAO inhibitors, and dopamine transporter inhibitors. Moreover, the present invention also relates to combinatorial libraries of polypharmacophoric compounds. Another aspect of the present invention relates to the use of a polypharmacophoric compound in a method of treating a mammal in need thereof. For example, a polypharmacophoric compound of the present invention may be used in a method of treating a mammal afflicted with Alzheimer&#39;s Disease, Huntington&#39;s Disease, depression, attention deficit disorder, autism, obesity, or inflammation.

RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalPatent Application Ser. No. 60/175,617, filed Jan. 11, 2000.

BACKGROUND OF THE INVENTION

The degeneration of 50-70% of the population of dopaminergic neurons inthe human brain results in the profound disturbances of motor functionthat are characteristic of Parkinson's Disease (PD). As thisdegeneration of neurons progresses, the symptoms become increasinglysevere, leading not only to loss of motor function but also to anincreased incidence of dementia and other neurological disorders.Currently, over a million people in North America are affected by thisdisease whose single most consistent risk factor is age. Because thepopulation of the elderly is expected to increase over the next fourdecades, it is projected that neurodegenerative diseases, such asParkinson's Disease, may pass cancer as the second most common cause ofdeath among the elderly. Therefore, the development of therapeuticagents that can delay the onset of disease, slow its progression, orenhance the effectiveness of other drugs, will provide a substantialcontribution to reducing the mortality and morbidity due to Parkinson'sDisease among the elderly and increasing the quality of life forafflicted individuals.

Treatment of Parkinson's Disease has traditionally been subdivided intothree categories: preventive, symptomatic and restorative intervention.The latter intervention, which include transplantation of adrenalmedulla cells, intraventricular delivery of dopaminergic neurotrophicfactor GDNF, and gene therapy, are in very early stages of safety andefficacy trials. Protective therapy with selective monoamine oxidase B(MAO-B) inhibitors, such as selegiline, has been unproductive to date.Early trials, which indicated promise, could largely be explainedthrough amelioration of symptoms rather than by slowing the progressionof the disease. As a result, most of the current efforts continue tofocus on therapeutic agents that affect symptoms which accompany thedisease rather than to reverse or prevent it. This remains an importantarea for medical research, due to the problems that exist with currenttherapeutic interventions.

Evaluation of the literature indicates that levodopa (L-DOPA) stillremains the agent of choice for the initial treatment of PD. There isclearly a beneficial motor response to this drug during the early statesof the disease, however, as the disease progresses, the effectiveness ofthe drug is reduced and other side effects become more pronounced.Patients may experiece fluctuations in motor response, dyskineasias, orpsychiatric disturbances, such as nightmares, hallucinations, psychosisor depression. Alternatives, for example, the use of amantidine,selegiline or anti-cholinergic agents may provide some initial benefit,but in most cases patients still require levodopa or other dopamine (DA)agonists for effective symptomatic relief. Even the DA agonists, whenused as monotherapeutic agents, often fail to exceed the effectivenessof levodopa.

The declining efficacy of the major therapeutic agents and theappearance of other manifestations during the course of the diseasesuggest additional therapeutic strategies. Among the proposed directionsare new formulations of levodopa to improve the delivery of the drug tothe affected region of the brain, selective serotonin reuptakeinhibitors (SSRI's) and monoamine oxidase inhibitors (MAO I) to treatdepression in PD patients, dopamine transporter (DAT), and catecholO-methyl transferase (COMT) inhibitors to prolong the effects of DA, andDA receptor (D1) agonists to reduce dyskinesias. All of these approachesutilize separate discrete molecular entities to elicit the desiredresponse, either alone or in combination with other agents. As such,they are subject to limitations often associated with combinationtherapy, for example, noncompliance due to different dosing schedulesand drug-drug interactions.

As shown in FIG. 1, which depicts a model dopaminergic neuron, inaddition to having the processes for dopamine synthesis, this regioncontains the dopamine transporter (DAT) which is responsible for removalof dopamine from the synapse, catechol O-methyl transferase (COMT) andmonoamine oxidases (MAO-A and B) which are involved in DA metabolism,and DA receptors (D1, D2, D2, etc.) which mediate dopaminergicresponses. Thus, intervention at one site alone or selectively, as isthe common for traditional therapeutics, may only produce a partialresponse because of compensatory mechanisms mediated by the other sites.Because of this decreased response, often a greater dose must beutilized which often results in adverse side effects. In contrast, anintervention strategy which affects several sites at a dose level thatmay be ineffective if present solely, may produce an additive responsethat would be therapeutically beneficial, while reducing the possibilityfor adverse side effects. Although FIG. 1 depicts the dopaminergicneuron, it will be appreciated that other regions having multiplereceptor sites in close proximity may be involved in other diseases andconditions, and thus also may utilize this intervention strategy.

Clearly, because of the need to increase the efficacy and safety ofpharmaceuticals, it would be beneficial to develop pharmaceuticals whichcontain multiple pharmacophoric sites capable of interacting at multiplebiological sites, preferably for those biological sites which act inconcert, implicated in specific diseases and conditions and/or involvedin side effects of these diseases or conditions.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts a model dopaminergic neuron.

FIG. 2 depicts the modification of dopamine action by one embodiment ofan inventive polypharmacophoric agent.

FIG. 3 depicts the contrast between hybrid drugs currently utilized andthe inventive scaffolded polypharmacophores.

FIG. 4 depicts certain preferred pharmacophoric fragments.

FIG. 5 depicts certain preferred components for the synthesis ofscaffolded polypharmacophores.

FIG. 6 depicts certain preferred components to be utilized in acombinatorial synthesis.

FIG. 7 depicts the representative synthesis of certain preferred vinylboronic acids.

FIG. 8 depicts the representative synthesis of certain preferredaldehydes.

FIG. 9 depicts certain preferred scaffolded polypharmacophores.

FIG. 1O depicts certain preferred scaffolded polypharmacophores.

FIG. 11 depicts certain preferred scaffolded polypharmacophores.

FIG. 12 depicts certain preferred scaffolded polypharmacophores.

FIG. 13 depicts the percent specific [¹²⁵I]RTI-55 bound vs. log [drug].

FIG. 14 depicts the percent specific [¹²⁵I]RTI-55 bound vs. log [drug].

FIG. 15 depicts the percent specific [¹²⁵I]RTI-55 bound vs. log [drug].

FIG. 16 depicts the percent specific [¹²⁵I]RTI-55 bound vs. log [drug].

FIG. 17 depicts the percent specific [¹²⁵I]RTI-55 bound vs. log [drug].

SUMMARY OF THE INVENTION

The present invention recognizes that it is often desirable, in thetreatment, prevention, and diagnosis of a disease or condition, toutilize an agent that is able to interact at more than one biologicalsite. For example, this may include, but is not limited to, eliciting orinhibiting a biological response for a condition that implicates morethan one receptor site, preferably for those biological receptor sitesthat act in concert, or eliciting or inhibiting biological responses, inaddition to those involved in the particular condition, to treat sideeffects. Thus, in recognition of the need for, and the desirability ofthis approach, in one aspect, the present invention provides novelpolypharmacophoric scaffolds, libraries thereof, and methods for makingsaid scaffolds and libraries thereof.

In general, the polypharmacophoric scaffolds comprise a scaffold unithaving at least two pharmacophoric units appended thereto, whereby eachis capable of interacting at a biological site and/or eliciting orinhibiting a desired biological response. In certain preferredembodiments, the pharmacorphoric units are selected for their ability toelicit a response at two or more biological receptor sites thatpreferably act in concert and optionally are either physically orspatially close (e.g., DA agonist and DAT inhibitors) or can befunctionally connected (for example, via the polypharmacophoroic unit).In certain embodiments, the scaffold units and/or the pharmacophoricunits have one or more modifier units attached thereto, whereby thesemodifier units are slected to facilitate the delivery, synthesis,activation, absorption, solubility or detection (e.g., by the use offluorescent or radioactive moieties, or biotin, to name a few) of thescaffolded pharmacophoric units.

The novel scaffolded polypharmacophores can be depicted generally byformulas (I) and (IA):

As depicted in formula (I), S comprises a scaffold unit; P comprises apharmacophore, wherein x is greater than or equal to 2; M comprises amodifier unit, wherein y is greater than or equal to 0, whereby each oneof P and M, for each occurrence, is appended to said scaffold unit andsaid scaffold unit does not participate directly in the desiredpharmacological activity. In addition to modifier units being directlyappended to the scaffold, in certain other embodiments, as shown in(IA), additional modifier units (D), wherein a and b, for eachoccurrence of x or y, are each independently greater than or equal tozero, may also be directly attached to one or more pharmacophores (P)and/or to one or more existing modifier units (M) that are attached tothe scaffold.

In certain embodiments, each additional modifier unit can be linkedsequentially to either an existing modifier, M, or to a pharmacophore,P, to generate either of the appendages: S-M-D₁-D ₂-D₃ (etc.) orS-P-D₁-D₂-D₃ (etc.). In certain other embodiments, each additionalmodifier can be linked directly to the pharnacophore (P) or modifierunit (M) to generate either of the appendages:

In still other embodiments, each additional modifier can, in certaininstances, be linked directly to the pharmacophore or modifier unit, andin other instances also be linked sequentially to generate combinationsof appendages as shown for certain exemplary appendages below:

In a preferred embodiment, the scaffolded polypharmacophores, asdepicted in (I) and (IA), are utilized in diseases and/or conditionsthat implicate the dopaminergic neuron, and that include two or moresignalling pathways and/or modulators thereof. Thus, the inventivepolypharmacophores can be utilized to treat any neurological disorder.In particularly preferred embodiments, the pharmacophores utilized inthe present invention are selected to interact at a biological site inthe region of the dopaminergic neuron, wherein each of saidpharmacophores are preferably independently selected from the groupconsisting of D-1 agonist, D-2 agonist, D-3 agonist, D-4 agonist,irreversible MAO-inhibitors, reversible MAO-inhibitors, monoaminetransporter inhibitors, COMT-inhibitors, MAO-inhibitors, DA transporterinhibitors, 5HT inhibitors, NET inhibitors, and GABA inhibitors. It isalso particularly preferred that the scaffolded polypharmacophorecomprises two, and more preferably three, pharmacophores.

In other embodiments of the present invention, polypharmacophoricscaffolds and libraries thereof are provided as shown in Formulas (II)and (IIA).

As shown in Formula (II), at least two of A, B, or C comprise apharmacophore. In certain preferred embodiments, A, B, and C eachcomprise a pharmacophore. In other preferred embodiments, at least twoof A, B, or C comprise a pharmacophore and one of A, B, or C comprises amodifier unit. As shown in Formula (IIA), in certain other preferredembodiments, one or more additional modifier units may also be attachedone or more of A, B, or C. As discussed in more detail above for Formula(I), modifiers may be attached sequentially to one or more of A, B, orC; modifiers may each be directly attached to one or more of A, B, or C;or modifiers may be attached both sequentially and directly to one ormore of A, B, or C. In particularly preferred embodiments, thepharmacophores utilized in the present invention are selected tointeract at a biological site in the region of the dopaminergic neuron,wherein each of said pharmacophores are preferably independentlyselected from the group consisting of D-1 agonist, D-2 agonist, D-3agonist, D-4 agonist, irreversible MAO-inhibitors, reversibleMAO-inhibitors, monoamine transporter inhibitors, COMT-inhibitors,MAO-inhibitors, and DA transporter inhibitors.

In another exemplary embodiment of the present invention,polypharmnacophoric scaffolds and libraries thereof are provided asshown in formulas (III) and (IIIA):

wherein A, B, and C each comprise a desired pharmacophore or modifierunit. It is particularly preferred that two of A, B, or C comprises apharmacophore and one of A, B, or C comprises a modifier unit. As shownin (IIIA), each of A, B, or C may also have one or more additionalmodifier units, D, attached thereto. As discussed in more detail abovefor Formula (I), modifiers may be attached sequentially to one or moreof A, B, or C; modifiers may each be directly attached to one or more ofA, B, or C; or modifiers may be attached both sequentially and directlyto one or more of A, B, or C. In particularly preferred embodiments, thepharmacophores utilized in the present invention are selected tointeract at a biological site in the region of the dopaminergic neuron,wherein each of said pharmacophores are preferably independentlyselected from the group consisting of D-1 agonist, D-2 agonist, D-3agonist, D-4 agonist, irreversible MAO-inhibitors, reversibleMAO-inhibitors, monoamine transporter inhibitors, COMT-inhibitors,MAO-inhibitors, and DA transporter inhibitors.

The invention also provides a method for determining one or morebiological activities of an inventive polypharnacophore or library ofpolypharmacophores comprising contacting a scaffolded polypharmacophoreor library of scaffolded polypharmacophores having any one of formulas(I), (IA), (II), (IIA), (III), or (IIIA) to a biological target, anddetermining a statistically significant change in a biochemical activityrelative to the level of biochemical activity in the absence of ascaffolded polypharmacophore.

In another aspect, the present invention also provides a pharmaceuticalcomposition comprising a compound of any one of formulas (I), (IA),(II), (IIA), (III) or (IIIA) as described herein; or a pharmaceuticallyacceptable salt thereof; in combination with a pharmaceuticallyacceptable diluent or carrier.

In yet another aspect, the invention provides a method for the treatmentof disorders and or conditions implicating multiple receptor sites(e.g., more than one), preferably those that act in concert, in ananimal, comprising administering a pharmaceutically effective dose of acompound of any one of formulas (I), (IA), (II), (IIA), (III), or(IIIA), or a pharmaceutically acceptable salt thereof. In a particularlypreferred embodiment, the present invention provides a method for thetreatment of conditions in which the dopaminergic system is implicated,comprising administering a pharmaceutically effective dose of any one ofcompounds of formulas (I), (IA), (II), (IIA), (III), or (IIIA) or apharmaceutically acceptable salt thereof. In preferred embodiments, themethod is used to modulate the function of the dopaminergic system.

The invention also provides the use of a compound of any one of formulas(I), (IA), (II), (IIA), (III), or (IIIA); or a pharmaceuticallyacceptable salt thereof; to prepare a medicament useful for treating acondition which implicates biological systems which act in concert. Inpreferred embodiments, the medicament is used for treating a conditionin which the dopaminergic system is implicated, and preferably themedicament is used to modulate the function of the dopaminergic system.

In yet another aspect, the present invention provides a compositioncomprising a compound of any one of formulas (I), (IA), (II), (IIA),(III), or (IIIA) or a pharmaceutically acceptable salt thereof, for usein medical therapy or diagnosis. In a preferred embodiment, theinvention provides a labeled compound comprising a radionuclide,fluroescent tag or other label or identifier, and a compound of any oneof formulas (I), (IA), (IA), (IIA), (III), or (IIIA), wherein any one ormore of the pharmacophoric units or modifier units are labeled, usingradiolabels, fluroescence or otherwise; or a pharmaceutically acceptablesalt thereof, as well as methods for using such labeled compounds as animaging or diagnostic agent (e.g., to identify, or evaluate the functionof, specific binding sites in a particular organ of interest).

Definitions

“Pharmacophore”: The term “pharmacophore”, as used herein, refers to anagent capable of having a biological effect.

“Modifier Unit”: The term “modifier unit”, as used herein, refers to anymoiety or combination of moieites capable of facilitating the delivery,synthesis, activation, solubility or other desirable property of aninventive scaffolded polypharmacophore. Exemplary modifier unitsinclude, but are not limited to, spacers, scaffold assemblers,bioactivating groups, and targeting agents, to name a few.

“Linker unit”: The term “linker unit”, as used herein, refers to amolecule, or group of molecules, connecting a solid support and acombinatorial library member. The linker may be comprised of a singlelinking molecule, or may comprise a linking molecule and a spacermolecule.

“Identifier Tag”: The term “identifier tag” as used herein, refers to ameans for recording a step in a series of reactions used in thesynthesis of a chemical library. For the purposes of this application,the terms encoded chemical library and tagged chemical library bothrefer to libraries containing a means for recording each step in thereaction sequence for the synthesis of the chemical library.

“Targeting Moiety”: The term “targeting moiety”, as used herein, refersto any molecular structure which assists one or more of the appendedpharmacophores or modifier units in localizing to a particular targetingarea, entering a target cell(s), and/or binding to a target receptor.For examples, lipids (including cationic, neutral, and steroidal lipids,virosomes, and liposomes), antibodies, lectins, ligands, sugars,steroids, hormones, nutrients and proteins can serve as targetingmoieties.

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups, branched-chain alkyl groups,cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, andcycloalkyl substituted alkyl groups. In preferred embodiments, astraight chain or branched chain alkyl has 30 or fewer carbon atoms inits backbone (e.g., C₁-C₃₀ for straight chain, C₃-C₃₀ for branchedchain), and more preferably 20 or fewer. Likewise, preferred cycloalkylshave from 4-10 carbon atoms in their ring structure, and more preferablyhave 5, 6 or 7 carbons in the ring structure.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification and claims is intended to include both “unsubstitutedalkyls” and “substituted alkyls”, the latter of which refers to alkylmoieties having substituents replacing a hydrogen on one or more carbonsof the hydrocarbon backbone. Such substituents can include, for example,a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an ester, aformate, or a ketone), a thiocarbonyl (such as a thioester, athioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphonate,a phosphinate, anamino, an amido, an amidine, an imine, a cyano, anitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate,sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or anaromatic or heteroaromatic moiety. It will be understood by thoseskilled in the art that the moieties substituted on the hydrocarbonchain can themselves be substituted, if appropriate. For instance, thesubstituents of a substituted alkyl may include substituted andunsubstituted forms of aminos, azidos, iminos, amidos, phosphoryls(including phosphonates and phosphinates), sulfonyls (includingsulfates, sulfonamidos, sulfamoyls and sulfonates), and silyl groups, aswell as ethers, alkylthios, carbonyls (including ketones, aldehydes,carboxylates, and esters), —CF₃, —CN and the like. Exemplary substitutedalkyls are described below. Cycloalkyls can be further substituted withalkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substitutedalkyls, —CF₃, —CN, and the like. The term “arylkyl”, as used herein,refers to an alkyl group substituted with an aryl group (e.g., anaromatic or heteroaromatic group).

The terms “alkenyl” and “alkynyl” refer to unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double or triple bond respectively.

The term “aryl” as used herein includes 5-, 6- and 7-memberedsingle-ring aromatic groups that may include from zero to fourheteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazineand pyrimidine, and the like. Those aryl groups having heteroatoms inthe ring structure may also be referred to as “aryl heterocycles” or“heteroaromatics”. The aromatic ring can be substituted at one or morering positions with such substituents as described above, as forexample, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,hydroxyl, amino, nitro, sulfhlydryl, imino, amido, phosphonate,phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,sulfonamido, ketone, aldehyde, ester, a heterocyclyl, an aromatic orheteroaromatic moiety, —CF₃, —CN, or the like.

The terms “heterocyclyl” or “heterocyclic group” refer to 4- to10-membered ringtructures, more preferably 4- to 7-membered rings, whichring structures include one to four heteroatoms. Heterocyclyl groupsinclude, for example, pyrrolidine, oxolane, thiolane, imidazole,oxazole, piperidine, piperazine, morpholine, lactones, lactams such asazetidinones and pyrrolidinones, sultams, sultones, and the like. Theheterocyclic ring can be substituted at one or more positions with suchsubstituents as described above, as for example, halogen, alkyl,aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro,sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl,silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, aheterocyclyl, an aromatic or heteroaromatic moiety, —CF₃, —CN, or thelike.

The terms “polycyclyl” or “polycyclic group” refer to two or more cyclicrings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/orheterocyclyls) in which two or more carbons are common to two adjoiningrings, e.g., the rings are “fused rings”. Rings that are joined throughnon-adjacent atoms are termed “bridged” rings. Each of the rings of thepolycycle can be substituted with such substituents as described above,as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate,phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromaticmoiety, —CF₃, —CN, or the like.

It will be noted that the structure of some of the compounds of thisinvention includes asymmetric carbon atoms. It is to be understoodaccordingly that the isomers arising from such asymmetry are includedwithin the scope of this invention. Such isomers are obtained insubstantially pure form by classical separation techniques and bysterically controlled synthesis.

The phrase “protecting group” as used herein, refers to a chemical groupthat reacts selectively with a desired functionality in good yield togive a derivative that is stable to further reactions for whichprotection is desired, can be selectively removed from the particularfunctionality that it protects to yield the desired functionality, andis removable in good yield by reagents compatible with the otherfunctional group(s) generated during the reactions. Examples of suchprotecting groups include esters of carboxylic acids, ethers of alcoholsand acetals and ketals of aldehydes and ketones.

It will be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance withpermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc.

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and nonaromaticsubstituents of organic compounds. Illustrative substituents include,for example, those described hereinabove. The permissible substituentscan be one or more and the same or different for appropriate organiccompounds. For purposes of this invention, the heteroatoms such asnitrogen may have hydrogen substituents and/or any permissiblesubstituents of organic compounds described herein which satisfy thevalencies of the heteroatoms. This invention is not intended to belimited in any manner by the permissible substituents of organiccompounds.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Alsofor purposes of this invention, the term “hydrocarbon” is contemplatedto include all permissible compounds having at least one hydrogen andone carbon atom. In a broad aspect, the permissible hydrocarbons includeacyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic organic compounds which can besubstituted or unsubstituted.

The term “solid support” refers to a material having a rigid orsemi-rigid surface. Such materials will preferably take the form ofsmall beads, pellets, disks, chips, dishes, multi-well plates, wafers orthe like, although other forms may be used. In some embodiments, atleast one surface of the substrate will be substantially flat. The term“surface” refers to any generally two-dimensional structure on a solidsubstrate and may have steps, ridges, kinks, terraces, and the likewithout ceasing to be a surface.

The term “polymeric support”, as used herein, refers to a soluble orinsoluble polymer to which an amino acid or other chemical moiety can becovalently bonded by reaction with a functional group of the polymericsupport. Many suitable polymeric supports are known, and include solublepolymers such as polyethylene glycols or polyvinyl alcohols, as well asinsoluble polymers such as polystyrene resins. A suitable polymericsupport includes functional groups such as those described below. Apolymeric support is termed “soluble” if a polymer, or apolymer-supported compound, is soluble under the conditions employed.However, in general, a soluble polymer can be rendered insoluble underdefined conditions. Accordingly, a polymeric support can be solubleunder certain conditions and insoluble under other conditions.

The phrase “therapeutically-effective amount” as used herein means thatamount of a compound, material, or composition of the present inventionwhich is effective for producing some desired therapeutic effect in atleast a sub-population of cells in an animal and thereby blocking thebiological consequences of that event in the treated cells, at areasonable benefit/risk ratio applicable to any medical treatment.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically-acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, includingbut not limited to, a liquid or solid filler, diluent, excipient,solvent or encapsulating material, involved in carrying or transportingthe subject scaffolded polypharmacophores from one organ, or portion ofthe body, to another organ, or portion of the body.

“Subject”: The term “subject”, as used herein, refers to a human oranimal (e.g., rat, mouse, cow, pig, horse, sheep, monkey, cat, dog,goat, etc.)

“Amino acid” or “amino acid residue” refers to any of the naturallyoccurring amino acids, as well as synthetic analogs and derivativesthereof. In general the abbreviations used herein for designating theamino acids and the protective groups are based on recommendations ofthe IUPAC-TUB Commission on Biochemical Nomenclature (see Biochemistry(1972) 11:1726-1732). “Amino acid” refers to any of the naturallyoccurring amino acids, as well as synthetic analogs and derivativesthereof. .alpha.-Amino acids comprise a carbon atom to which is bondedan amino group, a carboxyl group, a hydrogen atom, and a distinctivegroup referred to as a “side chain”. The side chains of naturallyoccurring amino acids are well known in the art and include, forexample, hydrogen (e.g., as in glycine), alkyl (e.g., as in alanine,valine, leucine, isoleucine, proline), substituted alkyl (e.g., as inthreonine, serine, methionine, cysteine, aspartic acid, asparagine,glutamic acid, glutamine, arginine, and lysine), arylalkyl (e.g., as inphenylalanine and tryptophan), substituted arylalkyl (e.g., as intyrosine), and heteroarylalkyl (e.g., as in histidine). See, e.g.,Harper et al (1977) Review of Physiological Chemistry, 16th Ed., LangeMedical Publications, pp. 21-24. One of skill in the art will appreciatethat the term “amino acid” also includes .beta.-, .gamma.-, .delta.-,and .omega.-amino acids, and the like. As used herein, the twentyconventional amino acids and their abbreviations follow conventionalusage (see IMMUNOLOGY-A SYNTHESIS, 2nd Edition, E. S. Golub and D. R.Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991), which isincorporated herein by reference). Amino acid residues are abbreviatedas follows: Phenylalanine is Phe or F; Leucine is Leu or L; Isoleucineis Ile or I; Methionine is Met or M; Norleucine is Nle; Valine is Val orV; Serine is Ser or S; Proline is Pro or P; Threonine is Thr or T;Alanine is Ala or A; Tyrosine is Tyr or Y; Histidine is His or H;Glutamine is Gln or Q; Asparagine is Asn or N; Lysine is Lys or K;Aspartic Acid is Asp or D; Glutamic Acid is Glu or E; Cysteine is Cys orC; Tryptophan is Trp or W; Arginine is Arg or R; Glycine is Gly or G,and X is any amino acid.

The term “amino acid” or “amino acid residue” further includes analogs,derivatives and congeners of any specific amino acid referred to herein,as well as C-terminal or N-terminal protected amino acid derivatives(e.g. modified with an N-terminal or C-terminal protecting group). Forexample, the present invention contemplates the use of amino acidanalogs wherein a side chain is lengthened or shortened while stillproviding a carboxyl, amino or other reactive precursor functional groupfor cyclization, as well as amino acid analogs having variant sidechains with appropriate functional groups). For instance, the subjectcompound can include an amino acid analog such as, for example,cyanoalanine, canavanine, djenkolic acid, norleucine, 3-phosphoserine,homoserine, dihydroxy-phenylalanine, 5-hydroxytryptophan,1-methylhistidine, 3-methylhistidine, diaminopimelic acid, omithine, ordiaminobutyric acid. Other naturally occurring amino acid metabolites orprecursors having side chains which are suitable herein will berecognized by those skilled in the art and are included in the scope ofthe present invention.

It will also be appreciated that unnatural amino acids are within thescope of the present invention, as set forth in, for example, Williams(ed.), Synthesis of Optically Active .alpha.-Amino Acids, Pergamon Press(1989); Evans et al., J. Amer. Chem. Soc., 112:4011-4030 (1990); Pu etal., J. Amer. Chem. Soc., 56:1280-1283 (1991); Williams et al., J. Amer.Chem. Soc., 113:9276-9286 (1991); and all references cited therein.Examples of unconventional amino acids include: 4-hydroxyproline,O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine,5-hydroxylysine, and other similar amino acids and imino acids (e.g.,4-hydroxyproline). In the polypeptide notation used herein, theleft-hand direction is the amino terminal direction and the right-handdirection is the carboxy-terminal direction, in accordance with standardusage and convention.

Also included are the (D) and (L) stereoisomers of such amino acids, orunnatural amino acids, when the structure of the amino acid admits ofstereoisomeric forms. The configuration of the amino acids and aminoacid residues herein are designated by the appropriate symbols (D), (L)or (DL), furthermore when the configuration is not designated the aminoacid or residue can have the configuration (D), (L) or (DL). It will benoted that the structure of some of the compounds of this inventionincludes asymmetric carbon atoms. It is to be understood accordinglythat the isomers arising from such asymmetry are included within thescope of this invention. Such isomers can be obtained in substantiallypure form by classical separation techniques and by stericallycontrolled synthesis. For the purposes of this application, unlessexpressly noted to the contrary, a named amino acid shall be construedto include both the (D) or (L) stereoisomers. D- and L-α-Amino acids arerepresented by the following Fischer projections and wedge-and-dashdrawings. In the majority of cases, D- and L-amino acids have R- andS-absolute configurations, respectively.

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and traiis-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

If, for instance, a particular enantiomer of a compound of the presentinvention is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.

Contemplated equivalents of the compounds described above includecompounds which otherwise correspond thereto, and which have the samegeneral properties thereof (e.g. the ability to bind to opioidreceptors), wherein one or more simple variations of substituents aremade which do not adversely affect the efficacy of the compound inbinding to opioid receptors. In general, the compounds of the presentinvention may be prepared by the methods illustrated in the generalreaction schemes as, for example, described below, or by modificationsthereof, using readily available starting materials, reagents andconventional synthesis procedures. In these reactions, it is alsopossible to make use of variants which are in themselves known, but arenot mentioned here.

DEATILED DESCRIPTION OF THE INVENTION

The present invention recognizes that it is often desirable, in thetreatment, prevention, and diagnosis of a disease or condition, toutilize an agent that is able to interact at more than one biologicalsite. For example, this may include, but is not limited to, eliciting orinhibiting a biological response for a condition that implicates morethan one receptor site (preferably for biological sites capable ofacting in concert), or eliciting or inhibiting biological responses, inaddition to those involved in the particular condition, to treat sideeffects. Thus, in recognition of the need for, and the desirability ofthis approach, in one aspect, the present invention provides novelpolypharmnacophoric scaffolds, libraries thereof, and methods for makingsaid scaffolds and libraries thereof.

As described previously, currently utilized approaches to treatment ofspecific diseases or conditions generally involve the use of separatediscrete molecular entities to elicit the desired response, either aloneor in combination with other agents. In contrast to this traditionalapproach, FIG. 2 depicts the inventive approach to the development oftherapeutics, in which a drug having multiple biological targets whichact is concert, is capable of exerting multiple beneficial effects. Forexample, referring to FIG. 2, inhibition of DAT by pharmacophorecomponent P would potentiate the effects of endogenous DA by preventingits reuptake. The binding of pharmacophore component a to the DAreceptor (DAR) would supplement the effects of DA, and inhibition ofCOMT by pharmacophore component χ would retard the metabolism ofendogenous DA. Because it would not be necessary to exert maximaleffects at any one site, the likelihood of drug interactions or toxicityat other neuronal sites is reduced.

In general, the novel polypharmacophores of the present inventioncomprise a molecular scaffold, or “scaffold unit”, as used herein, towhich pharmacophoric groups associated with a particular therapy can beappended or inserted. These polypharmacophores preferably exerttherapeutic effects for a particular condition in which multiplereceptor sites involved in the condition are in close proximity. Becausethe novel scaffolded polypharmacophores have appended groups, incontrast to traditional therapeutics which have embedded groups, it ispossible to control the spatial orientation of the particular appendedpharamacophores to obtain the maximum interaction between apharmacophore and a biological target, and thus exert the maximal effectfor a specific condition. In certain embodiments, the scaffold unitsand/or the pharmacophoric units have one or more modifier units attachedthereto, whereby these modifier units are slected to facilitate thedelivery, synthesis, activation, absorption, solubility or detection(e.g., by the use of fluorescent or radioactive moieties, or biotin, toname a few) of the scaffolded pharmnacophoric units, of the scaffoldedpharmacophoric units. In particularly preferred embodiments, theconstruction of the polypharmacophoric scaffold is amenable tocombinatorial chemistry techniques and thus libraries of the inventivescaffolded polypharmacophores can be generated and tested for biologicalactivity. FIG. 3 depicts the contrast between traditional hybrid drugscurrently utilized which typically incorporate or integrate twopharmacophoric units within the core structure of the drug, and theinventive scaffolded polypharmacophores, as also depicted by FIG. 2,which are more likely to retain the activity of the individualpharmacophoric groups. As will be appreciated by one of ordinary skillin the art, an agent that is capable of interacting at one or more sitescould also be useful as a diagnostic tool or agent.

The inventive scaffolded polypharmacophores as depicted generally informulas (I) and (IA), libraries thereof, and methods for making thesescaffolded polypharmacophores are described in more detail below.Certain other preferred embodiments of these scaffolds and libraries arealso depicted in formulas (II), (IIA), (III) and (IIIA) and aredescribed in more detail herein. The discussion of these specificexamples, however, is not intended to limit the scope of the presentinvention.

Scaffolded Polypharmacophores of the Present Invention:

As described earlier, many diseases and conditions are believed toimplicate more than one biological site and thus the inventivescaffolded polypharmacophores can be tailored for a specific therapeuticeffect. Specific diseases and conditions encompassed by the presentinvention include, but are not limited to, Parkinson's Disease,Alzheimer's Disease, Huntington's Disease, depression, Attention DeficitDisorder (ADD), autism, obesity, inflammation, rheumatoid diseases,cardiovascular diseases, hypertension, cancer and diabetes, to name afew. In particularly preferred embodiments, the scaffoldedpolypharmacophores are utilized for conditions in which the dopaminergicneuron is implicated. Other receptors capable of being targeted by thenovel polypharmacophores include, but are not limited to serotoninreceptors, metabolic glutamate receptors for epilepsy, NMDA receptors,AMPA receptors, Kainate receptors, peptide receptors, and nAGR, mACAR,and AchE receptors.

The present invention provides novel scaffolded polypharmacophores asdepicted generally by formulas (I) and (IA):

As depicted in formula (I), S comprises a scaffold unit; P comprises apharmacophore, wherein x is greater than or equal to 2; M comprises amodifier unit, wherein y is greater than or equal to 0, whereby each oneof P and M, for each occurrence, is appended to said scaffold unit andsaid scaffold unit does not participate directly in the desiredpharmacological activity. In addition to modifier units being directlyappended to the scaffold, in certain other embodiments additionalmodifier units (D), wherein a and b, for each occurrence of x or y, areeach independently greater than or equal to zero, may also be directlyattached to one or more pharmacophores (P) and/or to one or moreexisting modifier units (M) that are attached to the scaffold togenerate the general structure as shown in formula (IA).

It will be appreciated that, in one embodiment, each additional modifierunit can be linked sequentially to either an existing modifier, M, or toa pharmacophore , P, to generate either of the appendages: S-M-D₁-D₂-D₃(etc.) or S-P-D₁-D₂-D₃ (etc.). In other embodiments, each additionalmodifier can be linked directly to the pharmacophore (P) or modifierunit (M) to generate either of the appendages:

In still other embodiments, each additional modifier can, in certaininstances, be linked directly to the pharmacophore or modifier unit, andin other instances also be linked sequentially to generate combinationsof appendages as shown for certain exemplary appendages below:

It will be appreciated that a variety of pharmacophoric and modifierunits can be appended to the scaffold structures to achieve a desiredpharmacological effect. In general, the pharmacophoric unit will beselected to have a desired biological effect associated with a specificcondition, and the modifier unit will be selected to facilitate thedelivery, detection, synthesis, activation or solubility of an inventivescaffolded polypharmacophore. In preferred embodiments, pharmacophoricunits are each independently selected from the group consisting of D-1agonist, D-2 agonist, D-3 agonist, D-4 agonist, irreversibleMAO-inhibitors, reversible MAO-inhibitors, monoamine transporterinhibitors, COMT-inhibitors, MAO-inhibitors, and DA transporterinhibitors.

In a preferred embodiment of the present invention, polypharmacophoricscaffolds and libraries thereof are provided as shown in Formulas (II)and (IIA).

As shown in Formula (II), at least two of A, B, or C comprise apharmacophore. In certain preferred embodiments, A, B, and C eachcomprise a pharmacophore. In other preferred embodiments, at least twoof A, B, or C comprise a pharmacophore and one of A, B, or C comprises amodifier unit. As shown in Formula (IIA), in certain other preferredembodiments, one or more additional modifier units (D), wherein a, b,and c, are each independently greater than or equal to zero, may also beattached to one or more of A, B, or C. As discussed in more detail abovefor Formula (I), modifiers (D) may be attached sequentially to one ormore of A, B, or C; modifiers may each be directly attached to one ormore of A, B, or C; or modifiers may be attached both sequentially anddirectly to one or more of A, B, or C. In certain preferred embodiments,the novel scaffolded polypharmacophores are utilized to treat conditionsin which the dopaminergic system is implicated and the pharmacophoricunits are selected from the group consisting of D-1 agonist, D-2agonist, D-3 agonist, D-4 agonist, irreversible MAO-inhibitors,reversible MAO-inhibitors, monoamine transporter inhibitors,COMT-inhibitors, MAO-inhibitors, and DA transporter inhibitors.

In another exemplary embodiment of the present invention,polypharmacophoric scaffolds and libraries thereof are provided as shownin formulas (III) and (IIIA):

wherein A, B, and C each comprise a desired pharmacophore or modifierunit. It is particularly preferred that two of A, B, or C comprises apharmacophore and one of A, B, or C comprises a modifier unit. As shownin (IIIA), each of A, B, or C may also have one or more additionalmodifier units, D, attached thereto, wherein a, b, and c are eachindependently greater than or equal to zero. As discussed in more detailabove for Formnula (I), modifiers may be attached sequentially to one ormore of A, B, or C; modifiers may each be directly attached to one ormore of A, B, or C; or modifiers may be attached both sequentially anddirectly to one or more of A, B, or C. In certain particularly preferredembodiments, the scaffolded polypharmacophores are utilized to treatconditions in which the dopaminergic neuron is implicated and thepharmacophoric units are preferably selected from the group consistingof D-1 agonist, D-2 agonist, D-3 agonist, D-4 agonist, irreversibleMAO-inhibitors, reversible MAO-inhibitors, monoamine transporterinhibitors, COMT-inhibitors, MAO-inhibitors, and DA transporterinhibitors

Preferred pharmacophores and modifiers for use in the compounds andlibraries of the present invention will be described in more detailbelow.

Pharmacophores

It will be appreciated that a variety of phanmacophoric units can beappended to the scaffold structures to achieve a desired pharmacologicaleffect. As discussed previously, the compounds and libraries of thepresent invention can be utilized for a variety of conditions and/ordiseases that implicate more than one desired site for biologicalactivity, preferably for those biological sites capable of acting inconcert. In general, the pharmacophoric units will be selected to have adesired biological effect associated with a particular condition and/ordisease, which may involve selection of certain functional groupmoieties and/or selection of a specific spatial or stereochemicalorientation, and will also be selected for certain features, such asspecific functional groups that allow facile synthesis of the scaffoldedpolypharmacophores. In certain embodiments, phanmacophoric units arealso selected to enable the modification of the pharmacophoric unit witha modifier unit, as will be described in more detail below.

As will be appreciated by one of ordinary skill in the art, thereexists, for many conditions and diseases, a large collection of data andliterature describing the pharmacological effects of specific drugs andagents. Instead of utilizing simply one agent to treat a disease orcondition, the present invention utilizes several phanmacophoricmoieties to treat a disease or condition. Thus, the challenge is torationally design the target molecules (phaymacophores) in a manner thatwill retain significant biological activity at each of the targetedsites. Thus, the novel scaffolded polypharmacophores of the presentinvention utilize known pharmacophores and utilize structure-activityrelationships available for specific target sites in the development ofthe most efficacious phanmacophores. In general, a variety ofphamacophoric units can be utilized in the present invention. Specificpharmacophoric moieties include, but are not limited to small organicmolecules, peptides, peptidomimetics, nucleotides, and carbohydrates andwill be selected based upon the individual condition to be treated andthe pharmcological profile.

In a preferred embodiment, small organic molecules are utilized in thepresent invention as phannacophoric units. The use of small organicmolecules may confer increased stability and cell permeability. In aparticularly preferred embodiment of the present invention, the subjectscaffolded polyphanmacophores are utilized to treat Parkinson's Disease.Thus, several classes of phannacophoric units can be utilized forattachment to the scaffold including, but not limited to dopamineagonists, MAO-inhibitors, monoamine transporter inhibitors, and catecholO-methyl transferase inhibitors. FIG. 4 depicts, for A, B, C in Formula(III), preferred pharmacophoric fragments to be utilized for Parkinson'sDisease Therapeutics.

In yet another embodiment, the present invention utilizes amino acids,peptides, peptidomimetics or any combination of peptides orpeptidomimetics. A peptide for use in the inventive compounds andmethods comprises two or more amino acid residues. In general theabbreviations used herein for designating the amino acids and theprotective groups are based on recommendations of the IUPAC-IUBCommission on Biochemical Nomenclature (see Biochemistry (1972)11:1726-1732). In certain embodiments, the amino acids used in theapplication of this invention are those naturally occurring amino acidsfound in proteins, or the naturally occurring anabolic or catabolicproducts of such amino acids which contain amino and carboxyl groups.The term “amino acid” or “amino acid residue” further includes analogs,derivatives and congeners of any specific amino acid referred to herein,as well as C-terminal or N-terminal protected amino acid derivatives(e.g. modified with an N-terminal or C-terminal protecting group). Forexample, the present invention contemplates the use of amino acidanalogs wherein a side chain is lengthened or shortened while stillproviding a carboxyl, amino or other reactive precursor functional groupfor cyclization, as well as amino acid analogs having variant sidechains with appropriate functional groups). For instance, the subjectcompound can include an amino acid analog such as, for example,cyanoalanine, canavanine, djenkolic acid, norleucine, 3-phosphoserine,homoserine, dihydroxy-phenylalanine, 5-hydroxytryptophan,1-methylhistidine, 3-methylhistidine, diaminopimelic acid, omithine, ordiaminobutyric acid. Other naturally occurring amino acid metabolites orprecursors having side chains which are suitable herein will berecognized by those skilled in the art and are included in the scope ofthe present invention.

It will also be appreciated that unnatural amino acids are within thescope of the present invention, as set forth in, for example, Williams(ed.), Synthesis of Optically Active alpha.-Amino Acids, Pergamon Press(1989); Evans et al., J. Amer. Chem. Soc., 112:4011-4030 (1990); Pu etal., J. Amer. Chem. Soc., 56:1280-1283 (1991); Williams et al., J. Amer.Chem. Soc., 113:9276-9286 (1991); and all references cited therein.Examples of unconventional amino acids include: 4-hydroxyproline,O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine,5-hydroxylysine, and other similar amino acids and imino acids (e.g.,4-hydroxyproline).

Moreover, as will be appreciated by one of ordinary skill in the art, ifa desired peptide having a specific activity if found to be unsuitablefor various reasons such as chemical instability, or lack of cellpermeability, a suitable peptidomimetic can instead be utilized. Suchpeptidomimetics may comprise close analogs of the original peptideselected as a pharinacophoric unit, or such peptidomimetics may departfrom an original pharmacophoric peptide, and this incorporate only a fewor none of the original peptide features. Additionally, novel and random“peptide-like” or peptidomimetic compounds may be generated for use aspharmacophoric units to determine those compounds that may have a moredesirable activity. Such peptidomimetics can have such attributes asbeing non-hydrolyzable (e.g., increased stability against proteases orother physiological conditions which degrade the corresponding peptide),increased specificity and/or potency, and increased cell permeabilityfor intracellular localization of the peptidomimetic. For illustrativepurposes, peptide analogs of the present invention can be generatedusing, for example, benzodiazepines (e.g., see Freidinger et al. inPeptides: Chemistry and Biology, G. R. Marshall ed., ESCOM Publisher:Leiden, Netherlands, 1988), substituted gama lactam rings (Garvey et al.in Peptides: Chemistry and Biology, G. R. Marshall ed., ESCOM Publisher:Leiden, Netherlands, 1988, p 123), C-7 mimics (Huffman et al. inPeptides: Chemistry and Biology, G. R. Marshall ed., ESCOM Publisher:Leiden, Netherlands, 1988, p. 105), keto-methylene pseudopeptides(Ewenson et al. (1986) J Med Chem 29:295; and Ewenson et al. inPeptides: Structure and Function (Proceedings of the 9th AmericanPeptide Symposium) Pierce Chemical Co. Rockland, Ill., 1985), β-turndipeptide cores (Nagai et al. (1985) Tetrahedron Lett 26:647; and Satoet al. (1986) J Chem Soc Perkin Trans 1:1231), α-aminoalcohols (Gordonet al. (1985) Biochem Biophys Res Commun 126:419; and Dann et al. (1986)Biochem Biophys Res Communi 134:71), diaminoketones (Natarajan et al.(1984) Biochem Biophys Res Commun 124:141), and methyleneamino-modifed(Roark et al. in Peptides: Chemistry and Biology, G. R. Marshall ed.,ESCOM Publisher: Leiden, Netherlands, 1988, p134). Also, see generally,Session III: Analytic and synthetic methods, in in Peptides: Chemistryand Biology, G. R. Marshall ed., ESCOM Publisher: Leiden, Netherlands,1988)

In addition to a variety of sidechain replacements, the presentinvention specifically contemplates the use of conformationallyrestrained mimics of peptide secondary structure. Numerous surrogateshave been developed for the amide bond of peptides. Frequently exploitedsurrogates for the amide bond include the following groups (i)trans-olefins, (ii) fluoroalkene, (iii) methyleneamino, (iv)phosphonamides, and (v) sulfonamides.

Additionally, peptidomimietics based on more substantial modifications adesired peptide can be used. Peptidomimetics which fall in this categoryinclude (i) retro-inverso analogs, and (ii) N-alkyl glycine analogs(so-called peptoids).

Furthermore, the methods of combinatorial chemistry are being brought tobear, e,g,, by G. L. Verdine at Harvard University, on the developmentof new peptidomimetics (see,http://glviris.harvard.edu/frame_research.htm). For example, oneembodiment of a so-called “peptide morphing” strategy focuses on therandom generation of a library of peptide analogs that comprise a widerange of peptide bond substitutes.

Many other peptidomimetic structures are known in the art and can bereadily adapted for use in the present invention. To illustrate, a“peptidomimetic pharmacophore” may incorporate the1-azabicyclo[4.3.0]nonane surrogate (see Kim et al. (1997) J. Org. Chem.62:2847), or an N-acyl piperazic acid (see Xi et al. (1998) J. Am. Chem.Soc. 120:80), or a 2-substituted piperazine moiety as a constrainedamino acid analogue (see Williams et al. (1996) J. Med. Chem.39:1345-1348). In still other embodiments, certain amino acid residuescan be replaced with aryl and bi-aryl moieties, e.g., monocyclic orbicyclic aromatic or heteroaromatic nucleus, or a biaromatic,aromatic-heteroaromatic, or biheteroaromatic nucleus.

Modifier Units and Additional Modifier Units

Additionally, as described above, certain embodiments of the presentinvention include one or more modifier units that may be either attacheddirectly to the scaffold unit or may be attached to existing modifierunits and/or pharmacophoric units. For example, modifier units maycomprise spacers, scaffold assemblers, delivery modulators,bioactivating groups (that is, they can provide cleavage sites byesterases, enzyme substrate cleavage sites, or pH labile cleavage sitesto name a few) and targeting agents, including but not limited tobiotin/avidin, biotin, folates, and peptide receptors. Furthermore,modifier units may comprise solid support units in which a linking unitcomprising a linker and optionally a spacer can be used to attach theinventive scaffolded polypharmacophore to a solid support.

In one particularly preferred embodiment, a modifier unit, which can beattached directly to the scaffold, or can be attached to pharmacophoresor other modifier units, is a targeting moiety. A targeting moiety, asused herein, refers to any molecular structure which assists theinventive scaffolded polypharmacophore in localizing to a particulartarget area, entering a target cell(s), and/or binding to a targetreceptor. For example, lipids (including cationic, neutral, andsteroidal lipids, virosomes, and liposomes), antibodies, lectins,ligands, sugars, steroids, hormones, nutrients and proteins can serve astargeting moieties.

The targeting moiety, which assists the construct in localizing to aparticular target area, entering a target cell(s), and/or binding to atarget receptor, may be selected on the basis of the particularcondition or site to be treated. The targeting moiety may furthercomprise any of a number of different chemical entities. In oneembodiment, the targeting moiety is a small molecule.

A particularly preferred targeting moiety for use in the presentinvention is biotin, a naturally occurring vitamin, which has been shownto localize effectively to tumors and sites of infection. Furthermore,as described in U.S. Pat. No. 5,716,594, imaging agents and therapeuticshave been successfully delivered to such sites when coupled to biotin.Another preferred small molecule targeting moiety is folate (see U.S.Pat. No. 5,820,847). Folates are particularly useful in targeting cancercells, since a variety of carcinomas overexpress folate receptors. SeeLadino et al. (Int. J. Cancer 1997, 73(6): 859-6). Riboflavin and itsderivatives are other small molecule targeting moieties for targetingdelivery of constructs to cancer cells (see, for example, U.S. Pat. No.5,688,488). Additional nutrients believed to trigger receptor-mediatedendocytosis and therefore useful as targeting moieties include camitine,inositol, lipoic acid, niacin, pantothenic acid, thiamin, pyridoxal,ascorbic acid, and the lipid soluble vitamins A, D, E, and K. A secondexemplary type of small molecule targeting moiety includes steroidallipids, such as cholesterol, and steroidal hormones, such as estradiol,testosterone, etc.

In another embodiment, the targeting moiety may comprise a protein.Particular types of proteins may be selected based on knowncharacteristics of the target site or target cells. For example, theprobe can be an antibody either monoclonal or polyclonal, where acorresponding antigen is displayed at the target site. As a secondexample, certain cells, such as malignant cells and blood cells displayparticular carbohydroates, for which a corresponding lectin may serve asa targeting moiety. In situations wherein a certain receptor isexpressed by the target cells, the targeting moiety may comprise aprotein or peptidomimetic ligand capable of binding to that receptor.Proteins corresponding to known cell surface receptors (including lowdensity lipoproteins, transferrin and insulin), fibrinolytic enzymes,anti-HER2, platelet binding proteins such as annexins, and biologicalresponse modifiers (including interleukin, interferon, erythropoietinand colony-stimulating factor) are examples of preferred targetingmoieties. Also, anti-EGF receptor antibodies, which internalizefollowing binding to the receptor and traffic to the nucleus to anextent, are preferred targeting moieties for use in the presentinvention to facilitate delivery of Auger emitters and nucleus bindingdrugs to target cell nuclei.

A number of monoclonal antibodies that bind to a specific type of cellhave been developed, including monoclonal antibodies specific fortumor-associated antigens in humans. Among the many such monoclonalantibodies that may be used are anti-TAC, or other interleukin-2receptor antibodies; 9.2.27 and NR-ML-05 to the 250 kilodalton humanmelanoma-associated proteoglycan; and NR-LU-10 to a pancarcinomaglycoprotein. An antibody employed in the present invention may be anintact (whole) molecule, a fragment thereof, or a functional equivalentthereof. Examples of antibody fragments are F(ab′)₂, Fab′, Fab, sF_(v),F_(V) fragments, and minibodies, which may be produced by conventionalmethods or by genetic or protein engineering.

Other preferred targeting moieties include sugars (e.g., glucose,fucose, galactose, mannose) that are recognized by target-specificreceptors. For example, instant claimed constructs can be glycosylatedwith mannose residues (e.g., attached as C-glycosides to a freenitrogen) to yield targeted constructs having higher affinity binding totumors expressing mannose receptors (e.g., glioblastomas andgangliocytomas), and bacteria, which are also known to express mannosereceptors (Bertozzi, CR and MD Bednarski Carbohydrate Research 223:243(1992); J. Am. Chem. Soc. 114:2242,5543 (1992)), as well as potentiallyother infectious agents.

Additional ligands which may be suitable for use as targeting moietiesin the present invention include haptens, epitopes, and dsDNA fragmentsand analogs and derivatives thereof. Such moieties bind specifically toantibodies, fragments or analogs thereof, including mimetics (forhaptens and epitopes), and zinc finger proteins (for dsDNA fragments).

In yet another preferred embodiment, the modifier unit may comprise aspacer moiety or a solid support unit to enable the synthesis of theinventive scaffolded polypharmacophores using the techniques ofcombinatorial chemistry which will be discussed in more detail below.

Synthesis of Scaffolded Polypharmacophores

As discussed above, the development of the synthesis of the novelscaffolded polypharnacophores preferably take into consideration theease of synthesis and the ability to incorporate a variety ofpharmacophoric units. Thue, in general, the development of a novelscaffolded polypharmacophore for use in a specific treatment firstinvolves selecting specific desired pharmacophoric components, whereeach of said pharmacophoric components comprises a functionality capableof reacting with a functionality present on the other pharmacophoriccomponents; and reacting said components under conditions tosimultaneoulsly generate a scaffolded structure having saidpharmacophoric groups appended thereto.

In particularly preferred embodiments, inventive polypharmacophoricscaffolds are prepared by utilizing domino reactions in which a desirednumber of specific simple components or substrates is provided and uponreaction are capable, through sequences in which a bond formation (orbond-breaking process) is combined with the formation of a newfunctionality, which again forms a new bond and a new functionality andso on. Domino reactions have been reviewed in the art and a wide varietyof reactions can be employed to generate complex molecules in thisfashion (Tietze et al. Curr. Opin. Chem. Biol. 1998, 2, 363). Althoughthe use of domino reactions are preferred for the generation of thenovel scaffolded polypharmacophores, one of ordinary skill in the artwill realize that other reaction schemes may be utilized, although it ispreferable that these schemes are able to produce the desired compoundseasily and in good yield and are amenable to combinatorial techniques.

It will be appreciated that it is particularly preferred that each ofthe desired components may be modified so that they may be attached tothe solid support. The use of a solid support bound component isparticularly preferred because it enables the use of more rapid splitand pool techniques to generate larger libraries (e.g., greater than10,000 members) more easily.

A solid support, for the purposes of this invention, is defined as aninsoluble material to which compounds are attached during a synthesissequence. The use of a solid support is advantageous for the synthesisof libraries because the isolation of support-bound reaction productscan be accomplished simply by washing away reagents from thesupport-bound material and therefore the reaction can be driven tocompletion by the use of excess reagents. Additionally, the use of asolid support also enables the use of specific encoding techniques to“track” the identity of the inventive compounds in the library. A solidsupport can be any material which is an insoluble matrix and can have arigid or semi-rigid surface. Exemplary solid supports include, but arenot limited to, pellets, disks, capillaries, hollow fibers, needles,pins, solid fibers, cellulose beads, pore-glass beads, silica gels,polystyrene beads optionally cross-linked with divinylbenzene, graftedco-poly beads, poly-acrylamide beads, latex beads, dimethylacrylamidebeads optionally crosslinked with N-N′-bis-acryloylethylenediamine, andglass particles coated with a hydrophobic polymer. One of ordinary skillin the art will realize that the choice of particular solid support willbe limited by the compatability of the support with the reactionchemistry being utilized. In one particularly preferred embodiment, aTentagel amino resin, a composite of 1) a polystyrene bead crosslinkedwith divinylbenzene and 2) PEG (polyethylene glycol), is employed foruse in the present invention. Tentagel is a particularly useful solidsupport because it provides a versatile support for use in on-bead oroff-bead assays, and it also undergoes excellent swelling in solventsranging from toluene to water.

The compounds of the present invention may be attached directly to thesolid support or may be attached to the solid support through a linkingreagent. Direct attachment to the solid support may be useful if it isdesired not to detach the library member from the solid support. Forexample, for direct on-bead analysis of biological/pharmacologicalactivity or analysis of the compound structure, a stronger interactionbetween the library member and the solid support may be desirable.Alternatively, the use of a linking reagent may be useful if more facilecleavage of the inventive library members from the solid support isdesired.

Furthermore, any linking reagent used in the present invention maycomprise a single linking molecule, or alternatively may comprise alinking molecule and one or more spacer molecules. A spacer molecule isparticularly useful when the particular reaction conditions require thatthe linking molecule be separated from the library member, or ifadditional distance between the solid support/linking unit and thelibrary member is desired. In one particularly preferred embodiment,photocleavable linkers are employed to attach the solid phase resin tothe component. Photocleavable linkers are particularly advantageous forthe presently claimed invention because of the ability to use theselinkers in in vivo screening strategies. Once the inventive compound isreleased from the solid support via photocleavage, the inventivepolypharmacophore is able to enter the cell. Exemplary photocleavablelinkers include, but are not limited to ortho-Nitrobenzyl photolinkersand dithiane protected benzoin photolinkers. One of ordinary skill inthe art will realize that the method of the present invention is notlimited to the use of photocleavable linkers; rather other linkers maybe employed, preferably those that are capable of delivering the desiredcompounds in vivo.

Combinatorial Methods for the Synthesis ofPolypharmacophoric Libraries

According to the method of the present invention, the synthesis oflibraries from the above-described scaffold structures can be performedusing established combinatorial methods for solution phase, solid phase,or a combination of solution phase and solid phase synthesis techniques.The synthesis of combinatorial libraries is well known in the art andhas been reviewed (see, e.g., “Combinatorial Chemistry”, Chemical andEngineering News, Feb. 24, 1997, p. 43; Thompson, L. A., Ellman, J. A.,Chem. Rev. 1996, 96, 555.) One of ordinary skill in the art will realizethat the choice of method will depend upon the specific number ofcompounds to be synthesized, the specific reaction chemistry, and theavailability of specific instrumentation, such as roboticinstrumentation for the preparation and analysis of the inventivelibraries. In particularly preferred embodiments, the reactions to beperformed on the inventive scaffolds to generate the libraries areselected for their ability to proceed in high yield, and in astereoselective fashion, if applicable.

In one embodiment of the present invention, the inventive libraries aregenerated using a solution phase technique. Traditional advantages ofsolution phase techniques for the synthesis of combinatorial librariesinclude the availability of a much wider range of organic reactions, andthe relative ease with which products can be characterized. In apreferred embodiment, for the generation of a solution phasecombinatorial library, a parallel synthesis technique is utilized, inwhich all of the products are assembled separately in their own reactionvessels. In a particularly preferred parallel synthesis procedure, amicrotitre plate containing n rows and m columns of tiny wells which arecapable of holding a few milliliters of the solvent in which thereaction will occur, is utilized. It is possible to then use n variantsof reactant A, such as a carboxylic acid, and m variants of reactant B,such as an amide to obtain n×m variants, in n×m wells. One of ordinaryskill in the art will realize that this particular procedure is mostuseful when smaller libraries are desired, and the specific wells canprovide a ready means to identify the library members in a particularwell.

In another more particularly preferred embodiment of the presentinvention, a solid phase synthesis technique is utilized, in which thedesired scaffold structures are attached to the solid phase directly orthough a linking unit, as discussed above. Advantages of solid phasetechniques include the ability to more easily conduct multi-stepreactions and the ability to drive reactions to completion becauseexcess reagents can be utilized and the unreacted reagent washed away.Perhaps one of the most significant advantages of solid phase synthesisis the ability to use a technique called “split and pool”, in additionto the parallel synthesis technique, develped by Furka. (Furka et al.,Abstr. 14th Int. Congr. Biochem., Prague, Czechoslovakia, 1988, 5, 47;Furka et al., Int. J. Pept. Protein Res. 1991, 37, 487; Sebestyen etal., Bioorg. Med. Chem. Lett., 1993, 3, 413.) In this technique, amixture of related compounds can be made in the same reaction vessel,thus substantially reducing the number of containers required for thesynthesis of very large libraries, such as those containing as many asor more than one million library members. As an example, the solidsupport scaffolds can be divided into n vessels, where n represents thenumber species of reagent A to be reacted with the scaffold structures.After reaction, the contents from n vessels are combined and then splitinto m vessels, where m represents the number of species of reagent B tobe reacted with the scaffold structures. This procedure is repeateduntil the desired number of reagents is reacted with the scaffoldstructures to yield the inventive library.

The use of solid phase techniques in the present invention may alsoinclude the use of a specific encoding technique. Specific encodingtechniques have been reviewed by Czarnik. (Czamik, A. W., CurrentOpinion in Chemical Biology, 1997, 1, 60.) As used in the presentinvention, an encoding technique involves the use of a particular“identifiying agent” attached to the solid support, which enables thedetermination of the structure of a specific library member withoutreference to its spatial coordinates. One of ordinary skill in the artwill also realize that if smaller solid phase libraries are generated inspecific reaction wells, such as 96 well plates, or on plastic pins, thereaction history of these library members may also be identified bytheir spatial coordinates in the particular plate, and thus arespatially encoded. It is most preferred, however for large combinatoriallibraries, to use an alternative encoding technique to record thespecific reaction history.

Examples of particularly preferred alternative encoding techniques thatcan be utilized in the present invention include, but are not limitedto, spatial encoding techniques, graphical encoding techniques,including the “tea bag” method, chemical encoding methods, andspectrophotometric encoding methods. Spatial encoding refers torecording a reaction's history based on its location. Graphical encodingtechniques involve the coding of each synthesis platform to permit thegeneration of a relational database. Examples of preferredspectrophotometic encoding methods include the use of mass spectroscopy,fluorescence emission, and nuclear magnetic resonance spectroscopy. In amost preferred embodiment, chemical encoding methods are utilized, whichuses the structure of the reaction product to code for its identity.Decoding using this method can be performed on the solid phase or off ofthe solid phase. One of ordinary skill in the art will realize that theparticular encoding method to be used in the present invention must beselected based upon the number of library members desired, and thereaction chemistry employed.

In an exemplary embodiment of the method of the present invention, alibrary of at least 25 compounds is prepared, more preferably at least100 compounds and most preferably at least 500 compounds. Each of thereagents utilized are preferably selected for their ability to generatediversity and for their ability to react in high yield. For example, asdepicted in FIG. 6, a combinatorial synthesis may be conducted byselecting specific amine, aldehyde, and vinyl boronic acid components,and reacting these in a combinatorial fashion, using solution phase orsolid phase techniques. As one of ordinary skill in the art willrealize, the use also of a skip codon, or “blank”; at each step yieldsfurther diversity. Furthermore, in particularly preferred embodiments,if a solid phase technique is utilized, after each reaction step, thebeads are “tagged” to encode the particular reaction choice employed. Aswill be appreciated by one of ordinary skill in the art, although theuse of combinatorial techniques is preferably employed at the stage ofscaffold synthesis, combinatorial techniques can also be employed afterthe scaffolded polypharmacophore has been generated to add modifierunits and/or ftinctionalize pharmcophoric units. In but one example, ifa peptide pharmacophore is initially generated, this peptide maysubsequently be modified and/or lengthened as needed to optimize thepharmacophoric profile.

Subsequent characterization of the library members can be performedusing standard analytical techniques, such as mass spectrometry, NuclearMagnetic Resonance Spectroscopy, and gas chromatrograpy. One of ordinaryskill in the art will realize that the selection of a particularanalytical technique will depend upon whether the inventive librarymembers are in the solution phase or on the solid phase.

Biological Activity of Scaffolded Polypharmacophores

As discussed above, it would be desirable to identify scaffoldedpolypharmacophores of the present invention that are capable ofinteracting at a biological site, for example, modulating the biologicalactivity of a biological target, such as a protein, nucleic acid, lipidor combination thereof, whereby such identified compounds are useful inthe treatment and/or prevention of diseases or conditions, or are usefulas diagnostic agents.

In preferred embodiments, the compounds may be used in in vitro assays,or any other system that allows detection of a chemical or biologicalfunction. In general, according to the method of the present invention,one or more inventive scaffolded polypharmcophores is contacted with abiological target having a detectable biochemical activity. Suchbiological targets include, for example, enzymes, receptors, subunitsinvolved in the formation of multimeric complexes. Such multimericcomplex subunits may be characterized by catalytic capabilities (suchas, for example, an ability to catalyze substrate conversion), or mayalternatively be primarily active in binding to one or more othermolecules. The biological target can be provided in the form of apurified or semi-purified composition, a cell lysate, a whole cell ortissue, or even a whole organism. The level of biochemical activity isdetected in the presence of the compound and a statistically significantchange in the biochemical activity, relative to the level of biochemicalactivity in the absence of the compound, identifies the compound as amodulator, e.g., inhibitor or potentiator of the biological activity ofa target protein. In some cases, particularly where assays are done onwhole cells or organisms, the effect of the chemical compound may be toalter the amount, in addition to or instead of the activity, of theparticular biological target. “Modulators”, therefore, are chemicalcompounds that alter the level or activity of a particular target. Inparticularly preferred embodiments, multiple compounds of the inventivescaffolded polypharmacophoric libraries are assayed simultaneously in ahigh-throughput format, preferably allowing simultaneous analysis of atleast 25 compounds, preferably at least 100 compounds, and morepreferably at least 500 compounds.

Pharmaceutical Compositions

Once a specific desired effect on a biological target has beenassociated with a particular compound of the inventive library, one ormore of the compounds of the present invention may be utilized as atherapeutic agent for a particular medical condition. A therapeuticagent for use in the present invention may include any pharmacologicallyactive substances that produce a local or systemic effect in animals,preferably mammals, or humans. The term thus means any substanceintended for use in the diagnosis, cure, mitigation, treatment orprevention of disease or in the enhancement of desirable physical ormental development and conditions in an animal or human.

As will be appreciated by one of ordinary skill in the art, thetherapeutic agent may be administered orally, topically or via injectionby itself, or additionally may be provided as a pharmaceuticalcomposition comprising the therapeutic agent and a biologicallyacceptable carrier. The inventive compositions can be, but are notlimited to an aqueous solutions, emulsions, creams, ointments,suspensions, gels, and liposomal suspensions. Particularly preferredbiologically acceptable carriers include but are not limited to water,saline, Ringer's solution, dextrose solution and solutions of ethanol,glucose, sucrose, dextran, mannose, mannitol, sorbitol, polyethyleneglycol (PEG), phosphate, acetate, gelatin, collagen, Carbopol, andvegetable oils. It is also possible to include suitable preservatives,stabilizers, antioxidants, antimicrobials, and buffering agents, forexample including but not limited to BHA, BHT, citric acid, ascorbicacid, and tetracycline. The therapeutic agents of the presently claimedinvention may also be incorporated or encapsulated in a suitable polymermatrix or membrane, thus providing a sustained-release delivery devicesuitable for implantation near the site to be treated locally.

As one of ordinary skill in the art will realize, the amount of thetherapeutic agent required to treat any particular disorder will ofcourse vary depending upon the nature and severity of the disorder, theage and condition of the subject, and other factors readily determinedby one or ordinary skill in the art.

Other Uses:

It will be appreciated that the methods, compounds and libraries of thepresent invention can be utilized in various disciplines. For example,the scaffolded polypharmacophores of the present invention may also beused as imaging agents or diagnostic agents when labeled with aradionuclide, or fluorescent label. For example, a modifier unit maycomprise a radionuclide (such as tritium, iodine-125, iodine-131,iodine-123, iodine-124, astatine-210, carbon-61, carbon-14, nitrogen-13,fluorine-18) may be incorporated into, or attached directly to the corestructure, as by halogenation; or the radionuclide (such as Tc-99m,Re-186, Ga-68, Re-188, Y-90, Sm-153, Bi-212, Cu-67, Cu-64, and Cu-62, toname a few) may be attached to a linking group or bound by a chelatinggroup which is then attached to the compound directly, or by means of alinker. Radiolabeling techniques such as these are routinely used in theradiopharmaceutical industry.

Radiolabeled compounds of the invention are generally useful as imagingagents to diagnose neurological disease (e.g., a neurodegenerativedisease) or a mental condition or to follow the progression or treatmentof such a disease or condition in a mammal (e.g., a human). Theradiolabeled compounds of the invention can be conveniently used inconjunction with imaging techniques such as positron emission tomography(PET) or single photon emission computerized tomography (SPECT).

The inventive polypharmacophores may also be useful in the area ofmaterials science. Because of the reactive moieties present in thesecompounds, molecules such as lipids and other polymeric materials may beattached and thus generate potentially important biomaterials.

Equivalents

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to theinventive compositions and methods of use thereof described herein. Suchequivalents are considered to be within the scope of this invention andare covered by the following claims. Additionally, examples ofparticularly preferred embodiments are presented in the examples belowand are intended to more particularly describe the present invention,but are not intended to limit the scope of the presently claimedinvention.

Exemplification

Synthesis of Scaffolded Polypharmacophores

In but one example of the present invention, novel scaffoldedpolypharmacophores have been developed for use as agents for Parkinson'sDisease. These scaffolded polyphamiacophores, in contrast tomulti-component “cocktail” drugs, are expected to retain access to thecentral nervous system and have simplified pharmacokinetic properties.In a preferred embodiment, the scaffold itself can be assembled in asingle 3-component reaction (Petasis reaction) from relatively simplematerials that provide this approach with pharmacophoric diversity andthe ability to be extended to combinatorial expansion. FIG. 5 depictscertain preferred components for the synthesis of these scaffoldedpolypharmacophores. Scheme 1 below, depicts the simultaneous formationof an exemplary polypharmacophoric scaffold of the present invention.Specific embodiments are described in more detail below, however, one ofordinary skill in the art will appreciate that all equivalents areintended to be encompassed.

1) Experimental Procedures for the Synthesis of inventive compounds

-   a. General Procedure for the Synthesis of α-vinyl-α-amino acids    (Compounds RNH-VII-92, RHN-Z-24, RNH-Z-26, RNH-Z-39, RNH-Z-44,    RNH-Z-58, RNH-Z-63, RNH-Z-68, ZHN-84, ZHN-92, ZHN-93, ZNH-94): To a    solution of glyoxylic acid hydrate (1.0 equivalents) in 3 mL    dichloromethane was added sequentially the amino (1.0 equivalent)    and the trans-vinylboronic acid (1.0 equivalent). The reaction was    stirred at ambient temperature for 2 hours, filtered and the    precipitate was washed with cold dichloromethane. The product was    purified by recrystallization from methanol-isopropanol or by column    chromatography or silica gel using dichloromethane-methanol as the    eluent.    -   1. Compound RNH-Z-24: Isolated yield, 35%; melting point        164-166° C.; NMR (300 MHz) (CD₃OD): 7.20-7.54 (m, 10H), 6.95 (d,        J=15 Hz, 1H), 6.26 (dd, J=15 Hz, J=10 Hz, 1H), 4.18 (d, J=9.6        Hz, 1H), 3.72 (dd, 2H), 3.1-3.2 (m, 2H), 2.8-2.95 (m, 1H),        2.0-2.1 (m, 3H); C, H, N: calc'd. C=78.5, H=7.17, N=4.36; obsd.        C=78.1, H=7.17, N=4.49    -   2. Compound RNH-Z-26: Yield=85%, melting point=181-182° C.; NMR        (300 MHz) (CD₃OD) 7.28-7.56 (m, 9H), 6.97 (d, J=16 Hz, 1H), 6.26        (dd, 1H), 4.25 (d, 1H), 3.9 (m, 1H), 3.2-3.65 (m, 4H), 2.38 (m,        2H), 2.0 (m, 2H); Anal. Calcd. C=67.8, H=5.9, N=3.8; Obsd.        C=66.0, H=6.1, N=3.9    -   3. Compound RNH-VII-92: Yield=85%; melting point=187-188 C.; NMR        (300 MHz) (CD₃OD) 6.9-7.55 (m, 11H), 6.25 (dd, 1H), 4.21 (d,        1H), 3.45 (m, 8H); Anal. Calcd. C=74.5, H=6.8, N=8.7; Obsd.        C=74.1, H=6.8, N=8.7-   b. General Procedure for the Synthesis of α-vinyl-α-amino alcohols    (Compounds RNH-Z-59, RNH-Z-61, RNH-Z-64, RNH-Z-70): To a solution of    glycoaldehyde dimer (0.5 equivalents) in 3 mL ethanol were added the    amino (1.0 equivalents) and the vinylboronic acid (1.0 equivalents).    The reaction was stirred at ambient temperature for 6-24 hours. The    reaction mixture was evaporated to dryness and the resulting solid    was purified by column chromatography on silica gel using    dichloromethane and methanol as the eluent.    -   1. Compound RNH-Z59: Yield=55%; melting point=96-98 C; NMR (300        MHz) (CD₃OD) 7.10-7.50 (m, 10H), 6.62 (d, 1H), 6.29 (dd, 1H),        3.80 (m, 2H), 2.5 (m, 2H), 1.9 (m, 5H)-   c. General Procedure for the Synthesis of α-vinyl-α-amino amides    (Compounds RNH-Z-28, RNH-Z-31) (A): To the stirred solution of the    α-vinyl-α-amino acid (1.0 equivalents) in 10 mL DMF were added    sequentially triethylamine (3 equivalents), 1-hydroxybenzothiazole    (1.1 equivalents), the amine (1.1 equivalents) and carbodiimide    coupling reagent (1.2 equivalents). The reaction was stirred at    ambient temperature for 6-24 hours. The reaction mixture was    partitioned between water and ethyl acetate. The organic layer was    collected and evaporated to dryness. The resultant residue was    purified by chromatography on silica gel using    dichloromethane-methanol as the eluent. (B) To the stirred solution    of the aminoaldehyde in ethanol (1.0 equivalents) was added    sequentially the amine (1.0 equivalent) and the vinylboronic acid    (1.0 equivalent). The reaction was allowed to proceed at ambient    temperature for 6-24 hours. The reaction mixture was evaporated to    dryness and the residue was purified by column chromatography using    dichloromethane-methanol as the eluent.    -   1. Compound RNH-Z-28: Yield=50%; NMR (300 MHz) (CD₃OD); 7.1-7.5        (m, 10H); 6.90 (d, 1H); 6.36 (dd, 1H); 4.04 (d, 1H); 3.0-3.4 (m,        10H), 2.5 (m, 1H), 2.2-2.4 (m, 2H); 1.8 (m, 4H)    -   2. Compound RNH-Z-31: Yield=60%; NMR (300 MHz) (CD₃OD); 7.1-8.1        (m, 11H); 6.67 (d, 1H); 6.26 (dd, 1H); 3.52 (d, 1H), 3.0-3.1 (m,        2H); 2.5 (m, 1H); 2.1-2.2 (m, 2H); 1.7 (m, 4H)        2) Generation of Combinatorial Libraries of Inventive        Polyphamcophores:

In an exemplary embodiment of the present invention, a directedcombinatorial library can be prepared for use as agents for treatingParkinson's Disease. The reactions and conditions described in the aboveexamples can also be utilized to generate these combinatorial librariesof polypharmacophores. These libraries can be prepared in the solutionphase or on the solid phase. We have selected for the amine component ofthe scaffold a D₂ agonist, an irreversible MAO inhibitor, and a DATinhibitor. The aldehyde and vinylboronic acid components incorporateinhibitors of COMT, MAO, and DAT. FIG. 6 depicts the use of certainexemplary fragments for a small combinatorial library for use as agentsfor Parkinson's Disease, although it will be appreciated that otherpharmacophores can also be utilized in the present invention. Although anumber of secondary amines, aldehydes and vinyl boronic acids shown inFIG. 6 are commercially available, many of the starting materialsrequire synthesis from other intermediates. Representative syntheses ofcertain components used in the inventive library are described in FIGS.7, and 8.

The synthesis of the mini-directed library is preferably performed on ascale which will provide approximately 0.1-1.0 mmoles of each targetcompound. Using the reaction conditions described above, solutions ofthe 3 representative amines, 3 aldehydes and 3 vinylboronic acids for3×3×3 library are prepared. The solvent is removed by rotary evaporationand each of the twenty seven compounds are isolated by columnchromatography on silica gel and recrystallized as their maleate orfumarate salt. Each compound is also characterized by ¹H-NMR, FTIR andHRMS (or elemental analysis) to confirm its structural identity.Clearly, these syntheses will generate mixtures of enantiomers anddiastereomers, which can be separated using techniques well-known in theart of organic synthesis, if desired.

3) Biological Evaluation of Agents in Theraputically Relevant BiologicalAssays

To evaluate the potential efficacy of the target compounds and librariesof compounds, in vitro screening assays are utilized against thebiological targets of the appended pharmacophores. For example,compounds that contain a dopamine receptor agonist, COMT inhibitor and aMAO inhibitor pharmacophore within their structure are evaluated inthose screens. The compounds will be compared to their separatecomponent to determine the extent to which the desired biologicalactivity is retained or enhanced. After this initial screening, thecompounds are then screened more extensively to determine their broaderpharmacological profile since this may identify additional benefits orpotential side effects.

Competitive binding assays are utilized to determine the affinity of theligands for the dopamine D-1 and D₂ receptors (human recombinantreceptor with [³H]SCH-23390 and [³H]spiperone as the radioligands).Efficacy of ligands which demonstrate K_(d) values <10⁷ are determinedin functional assays appropriate to the receptor subtype. Inhibition ofDAT binding (human recombinant protein in CHO cells) and [³H]DA reuptakein CHO-K1/hDAT cells that have been stabely transfected. The ability ofthe target ligands to inhibit COMT are determined using the enzymepreparation obtained from rats, specifically looking for the reductionin the metabolism of 3,4-dihydroxybenzoic acid to generate IC₅₀ values.Inhibition of MAO-B is determined in preparations obtained from ratliver using [³H]DA as the standard substrate to yield the IC₅₀ values.Selected compounds can also be more broadly screened as potentialreceptor ligands.

As depicted in Table 1, the drug effects on specific [¹²⁵I]RTI-55Binding to the Human Dopamine Transporter were examined. To characterizedrug interactions with the human dopamine transporter (hDAT), drugs(approximately 30 nM to 10 μM) were incubated with a membranepreparation from HEK-293 cells stably expressing the recombinant hDAT,[¹²⁵1I]RTI-55 (34 pM), and buffer in a final volume of 250 μl.Independent assays were conducted 2 to 3 times with duplicatedeterminations. The Cheng-Prusoff equation was used to convert IC₅₀values to K_(i) values. Data in Table 1 represent averaged values +/−range or +/− the s.e.m. for 2 or 3 experiments, respectively. The valuefor GBR-12935 is taken from Eshleman et al., 1999. TABLE 1 Drug Effectson Specific [¹²⁵I]RTI-55 Binding to the Human Dopamine Transporter DrugK_(i) value (nM) Hill Coefficient ZHN-84 75 ± 39 −0.5 ± 0.2 ZHN-92 3713± 1247 −1.2 ± 0.2 ZHN-93 727 ± 291 −1.0 ± 0.3 ZHN-94 2784 ± 1250 −0.8 ±0.2 RNH-Z-28 3922 ± 1245 −2.1 ± 0.6 RNH-Z-31 7727 ± 3074 −7.0 ± 4.5RNH-Z-44 3300 ± 855  −2.2 ± 0.9 GBR-12935 14 ± 3  −1.6 ± 0.8

Additionally, FIGS. 13-17 depict the percent specific [¹²⁵I] RTI-55bound versus the log[drug].

Additionally, as shown in Table 2, the ability of specific compounds tobind to multiple receptors is examined. TABLE II [¹²⁵I]RTI-55[¹²⁵I]RTI-55 [¹²⁵I]RTI-55 Binding in [³H]DA Binding in [³H]5HT Bindingin [³H]NE HEK hDAT Uptake in HEK hSERT Uptake in HEK-hNET Uptake inMembranes HEK hDAT Membranes HEK-hSERT Membranes HEK-hNET K_(i)(nM) semcells IC₅₀ K_(i) (nM) sem cells IC₅₀ K_(I) (nM) sem cells IC₅₀ CompoundHill slope (nM) sem Hill slope (nM) sem Hill slope (nM) sem GBR-12935 73 ± 39 18 ± 3  2091 ± 756 3710 ± 1429 628 ± 59 165 ± 17 −0.44 ± 0.09−1.47 ± 0.36 −0.90 ± 0.07 ZHN-84  74 ± 39 89 ± 40 >10 μM >10 μM >10 μM 805 ± 206 −0.52 ± 0.17 ZHN-92  3713 1247 2013 ± 428  >10 μM >10 μM >10μM 113 ± 15 −1.15 0.15 ZHN-93  727 ± 291 182 ± 59  >10 μM >10 μM  4540 ±1539 117 ± 20 −1.02 ± 0.30 −1.78 ± 0.65 ZHN-94  2784 ± 1250 385 ±111 >10 μM >10 μM >10 μM 2195 ± 470 −0.78 ± 0.17 RNH-Z-24 >10 μM >10μM >10 μM >10 μM >10 μM 204 ± 85 RNH-Z-28  3922 ± 1245 4045 ± 1630 >10μM >10 μM >10 μM 211 ± 65 −2.09 ± 0.60 RNH-Z-31  7727 ± 3074 >10 μM >10μM >10 μM >10 μM >10 μM −7.00 ± 4.54 RNH-Z-39 >10 μM >10 μM >10 μM >10μM  2920 ± 1226 >10 μM −0.96 ± 0.17 RNH-Z-44 3300 ± 855  1820 ± 819 >10μM >10 μM >10 μM >10 μM −2.16 ± −0.89 RNH-Z-48 >10 μM >10 μM >10 μM >10μM >10 μM >10 μM RNH-Z-52 >10 μM >10 μM >10 μM >10 μM >10 μM >10 μMRNH-Z-58 >10 μM >10 μM >10 μM >10 μM >10 μM >10 μM RNH-Z-59 >10 μM 857 ±168 >10 μM >10 μM >10 μM >10 μM

As will be appreciated by one of ordinary skill in the art, the resultsfor each of the compounds evaluated can be compiled to build structureactivity relationships at each of the targeted sites. The profilecontains, in addition to the physicochemical and spectroscopicproperties of the compound, the biological activity of the material,e.g., K_(d), K_(i), IC₅₀ values, compared to a mono-pharmacophoricstandard. The structure activity relationships obtained from the initialstudies can serve to direct the synthesis of subsequent larger directedlibraries of compounds with improved properties and as the basis for theselecting candidate agents for pre-clinical evaluation. Although theexamples above are directed to receptor sites believed to be involved inParkinson's Disease, depression, substance addiction, ADD, ADHD,Huntington's Disease, and schizophrenia, it will be appreciated by oneof ordinary skill in the art that other assays can be utilized toexamine the biological effects of other polypharmacophoric compounds,that are targeted for different conditions. For example, otherconditions include, but are not limited to asthma, inflammation, CHF,and hypertension.

In certain preferred embodiments, properties that one would expect forthe hybrid pharmacophoric compounds is activity against two of thetarget sites equal to that of the isolated pharmacophoric unit andactivity against a third target site with a reduction of 0.5 log unitsin potency as compared to the unsubstituted fragment. In anotherpreferred embodiment, the desired properties for the hybridpharmacophoric compounds comprise three pharmacological activities thatare reduced by 1 log unit. In a particularly preferred embodiment,optimal properties that one would expect for the hybrid pharmacophoriccompounds comprises activity against each of the target sites equal tothat of the isolated pharmacophoric unit. For example, if the hybridcontains a dopamine agonist, DAT inhibitor and a COMT inhibitor, ascomponents, the intact molecule would preferably express all threeactivities in approximately the same potencies as seem for eachfragment. It will be appreciated that preferred polypharmacophoriccompounds, in addition to comprising desirable activity against each ofthe target sites, also have desirable safety profiles. Evaluation andinterpretation of the data enables the determination of which componentsof the hybrid would require enhancement or reduction. This informationwill also influence the synthetic strategy for desired combinatoriallibraries and compounds.

REFERENCES

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Each of the references cited above is incorporated by reference.

1-54. (canceled)
 55. A polypharmacophore selected from the groupconsisting of:


56. A polypharmacophore represented by formula (II):

wherein: R₁₀ is selected from the group consisting of phenyl andsubstituted phenyl; R₁₁ is selected from the group consisting of phenyland substituted phenyl; and X is selected from the group consisting ofCH and N.
 57. The polypharrnacophore of claim 56, wherein each of R₁₀and R₁₁ is phenyl.
 58. The polypharmnacophore of claim 57, wherein eachof R₁₀ and R₁₁ is substituted phenyl.
 59. The polypharmnacophore ofclaim 58, wherein said substituted phenyl is 4-fluorophenyl.
 60. Apharmaceutical composition comprising a polypharmacophore of claim 55 or56, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable diluent or carrier.